11-16-2016, 11:39 PM
Join Date: Dec 2015
Location: Third rock from the sun
Micro propagation (aka tissue culture)
I’ve been screwing around with tissue culture for awhile now and I read somewhere that scientist were able to get 1000 year old seeds to grow that were found in ancient tombs and I thought why not use that for old cannabis seeds. So I started working on some recipes to pop old seeds.
What tissue culture is mostly used for is growing parts of plants like leaves, stems, flowers, or growing tips, technically you only need one living cell or a hand full of cells to grow plants through “in vitro” to grow a whole plant. If you’re good enough you can grow 1000’s of clones from one leaf it just takes time, pretty cool!
With these seeds I didn’t have to go crazy with the TC because I knew the cells within the seeds were still good from my other tests, they were just lacking the hormones, enzymes, auxins that kick start the germination process. So I developed some TC recipes that would aid in that process. So what does a seed need to germinate? They need grow regulators which are auxins, cytokinin, gibberellins and hormones like GA3 (gibberellic acid), benzylaminopurine (BAP), naphthaleneacetic acid (NAA). They also need sugars, nutrients, heat and moisture.
Things you’ll need to mix up a batch of TC medium:
Air tight Containers that can handle a pressure cooker or autoclave.( I use canning jars).
Flow hood or homemade clean box
3% Hydrogen peroxide
20grams of Sugar
4 to 6 grams of Agar (TC medium)
2.1 grams of Murashige & Skoog nutrient formula
1/2ml of Benzylaminopurine (BAP)
1/2ml of Naphthaleneacetic acid (NAA)
1/2ml of Thiamine B1
1ml PPM (TC preservative)
Food coloring (optional)
PH pen or drops
PH up and down
Scale to weigh out ingredients
These are not that strong of a mix you might get better results with a stronger mix…
Recipe #1: (red mix)
1 quart distilled water, 4 grams agar, 1/2ml BAP 1/2ml NAA, 1/2ml B1, 2.1 grams M&S, 20 grams sugar, 1ml ppm, a few drops of red food coloring.
Recipe #2: (blue mix)
1 quart distilled water, 4 grams agar, 1/2ml B1, 2.1 grams M&S, 20 grams sugar, 1ml ppm, a few drops of blue food coloring.
Ok start by mixing the ingredients then heating the mixture just until the agar dissolves, let cool down and then PH the mix to 5.6 to 5.8 and pour into your jars, put lids on loose and pressure cook for 30 minute at 15psi, let everything cool down and your lids should pop down and you’ll know they are sealed. Put in refrigerator overnight. Next day get everything you’re going to need tweezers, alcohol, paper towels, clean box, hydrogen peroxide, etc.
Start by mixing 50/50 sterile water and 3% hydrogen peroxide (when pressure cooking your mix, put a jar or two of distilled water in the cooker too, that’s your sterile water) this is what you’ll be cleaning and sterilizing your seeds in for one hour, when their soaking clean all of the tools, hands and surfaces you’ll be working with in 90% alcohol, Once the seeds are all clean place them in the jar and in medium about a quarter inch down as quickly as you can to limit the chance of contamination. Now seal them up and put them on a heat mate set to 75F. You should see them sprout in a couple weeks or so, but it could take months so be patient.
I still have some time to kill on these; I just started them about 3 weeks ago but so far the red mix is popping more seeds I guess the added growth regulators helped out. I’m up to 20% germination rate and about 10% with the blue mix. Also two out of the five jars got contaminated so that messes up the numbers.
Last edited by Mystic Funk; 11-17-2016 at 05:04 AM..
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